Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Sci Rep. 2023 May 4;13(1):7249. doi: 10.1038/s41598-023-34289-3.
L-Lactate is a major waste compound in cultured animal cells. To develop a sustainable animal cell culture system, we aimed to study the consumption of L-lactate using a photosynthetic microorganism. As genes involved in L-lactate utilization were not found in most cyanobacteria and microalgae, we introduced the NAD-independent L-lactate dehydrogenase gene from Escherichia coli (lldD) into Synechococcus sp. PCC 7002. The lldD-expressing strain consumed L-lactate added to basal medium. This consumption was accelerated by expression of a lactate permease gene from E. coli (lldP) and an increase in culture temperature. Intracellular levels of acetyl-CoA, citrate, 2-oxoglutarate, succinate, and malate, and extracellular levels of 2-oxoglutarate, succinate, and malate, increased during L-lactate utilization, suggesting that the metabolic flux from L-lactate was distributed toward the tricarboxylic acid cycle. This study provides a perspective on L-lactate treatment by photosynthetic microorganisms, which would increase the feasibility of animal cell culture industries.
L-乳酸是培养的动物细胞中的主要废物化合物。为了开发可持续的动物细胞培养系统,我们旨在使用光合微生物研究 L-乳酸的消耗。由于大多数蓝藻和微藻中未发现与 L-乳酸利用相关的基因,我们将大肠杆菌(lldD)的 NAD 独立 L-乳酸脱氢酶基因引入到聚球藻 PCC 7002 中。表达 lldD 的菌株可以消耗添加到基础培养基中的 L-乳酸。通过表达大肠杆菌的乳酸透酶基因(lldP)和提高培养温度,可以加速 L-乳酸的消耗。在 L-乳酸利用过程中,细胞内乙酰辅酶 A、柠檬酸、2-酮戊二酸、琥珀酸和苹果酸的水平以及细胞外 2-酮戊二酸、琥珀酸和苹果酸的水平升高,表明 L-乳酸的代谢通量分布到三羧酸循环中。这项研究为光合微生物处理 L-乳酸提供了新视角,这将提高动物细胞培养行业的可行性。